Kyse 450

Human oesophageal epithelial cells (HET-1A) and ESCC cells (Eca109) were purchased from Fenghuishengwu (Wuhan, China), and ESCC cells (KYSE450, KYSE150 and KYSE30) were bought from COBIOER (Nanjing, China). The cell culture medium was the RPMI-1640 medium (Invitrogen, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (FBS; Invitrogen). All cells were maintained in a 5% CO₂ incubator at 37℃. Eca109 and KYSE150 cells were exposed to increasing doses of Taxol (Solarbio, Beijing, China) to establish Taxol-resistant ESCC cell lines (Eca109/Taxol and KYSE150/Taxol). Then, Taxol-resistant cells were treated with Taxol (10 nM) to maintain a resistant phenotype.
The small interfering RNA (siRNA) targeting circ_0006168 or JMJD1C (si-circ_0006168 or si-JMJD1C) and matched control (si-control), pcDNA-JMJD1C overexpression plasmid (pcDNA-JMJD1C) and matched control (pcDNA), miR-194-5p mimic (miR-194-5p) and matched control (miR-NC), miR-194-5p inhibitor (inhibitor-miR-194-5p) and matched control (inhibitor-NC) were obtained from Genecreat (Wuhan, China). Lentivirus-mediated short hairpin RNA (shRNA) targeting circ_0006168 (sh-circ_0006168) and corresponding control (sh-control) were provided by Genechem (Shanghai, China). Transient transfection was carried out in our research by using Lipofectamine 3000 Reagent (Invitrogen).

The normal esophageal epithelial cell line (NE3), originally developed at Hong Kong University, was obtained from Professor Li Fu (Department of Pharmacology, Shenzhen University). Other human ESCC cell lines, including Eca-109 (RRID: CVCL_6898), KYSE-30 (RRID: CVCL_1351), KYSE-70 (RRID: CVCL_1356), KYSE-180 (RRID: CVCL_1349), KYSE-450 (RRID: CVCL_1353), and TE1 (RRID: CVCL_1759) were purchased from the Cobioer Biosciences Co., Nanjing, China. NE3, Eca-109, KYSE30, KYSE70, KYSE180, KYSE450, and TE1 cells were maintained in 10% FCS-RPMI-1640 medium (Invitrogen, Shanghai, China). NE3 cells were cultured in EpiCM 4101 medium (ScienCell, USA). The cells were cultured in humidified atmosphere of 5% CO₂ at 37°C. All cell lines were authenticated by matching the short-tandem repeat (STR) DNA profiles of the cell to the corresponding standard STR in the database of ATCC (the American Type Culture Collection) and DSMZ (Deutsche Sammlung von Mikroorganismen und Zellkulturen). All cells were routinely tested and found negative for mycoplasma.